Genetic alphabet expansion transcription generating functional RNA molecules containing a five-letter alphabet including modified unnatural and natural base nucleotides by thermostable T7 RNA polymerase variants
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Genetic alphabet expansion transcription generating functional RNA molecules containing a five-letter alphabet including modified unnatural and natural base nucleotides by thermostable T7 RNA polymerase variants
Genetic alphabet expansion transcription generating functional RNA molecules containing a five-letter alphabet including modified unnatural and natural base nucleotides by thermostable T7 RNA polymerase variants
Thermostable T7 RNA polymerase variants were explored for genetic alphabet expansion transcription involving the unnatural Ds‒Pa pair. One variant exhibited high incorporation efficiencies of functionally modified Pa substrates and enabled the simultaneous incorporation of 2ʹ-fluoro-nucleoside triphosphates of pyrimidines into transcripts, allowing the generation of novel, highly functional RNA molecules.
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This work was supported by IBN (Biomedical Research Council, Agency for Science, A*STAR, Singapore) (to M.K. and I.H.), JST/PRESTO [JPMJPR13K9 to M.K.], NIH [5U54EB015403-02 to A.D.E.], Welch Foundation [F-1654 to A.D.E.], and National Security Science and Engineering Faculty Fellowship [FA9550-10-1-0169 to A.D.E.].